CN106472281A - A kind of liver moss is combined the breeding method of skinning with desert algae - Google Patents

A kind of liver moss is combined the breeding method of skinning with desert algae Download PDF

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CN106472281A
CN106472281A CN201610899794.9A CN201610899794A CN106472281A CN 106472281 A CN106472281 A CN 106472281A CN 201610899794 A CN201610899794 A CN 201610899794A CN 106472281 A CN106472281 A CN 106472281A
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moss
liver moss
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CN106472281B (en
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李敦海
彭成荣
郑娇莉
黄顺
张金丽
刘永定
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Institute of Hydrobiology of CAS
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G31/00—Soilless cultivation, e.g. hydroponics
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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    • A01G22/00—Cultivation of specific crops or plants not otherwise provided for
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
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    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/12—Unicellular algae; Culture media therefor

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Abstract

The invention discloses a kind of liver moss is combined the breeding method of skinning with desert algae, step is：A, the advantage liver moss species in summer collection area natural habitat to be repaired；B, the activation recovering of liver moss；C, with the moss crust after distilled water flushing activation recovering, obtain liver moss gametophyte；Liver moss gametophyte after D, sterilization is put in solid medium carries out culture acquisition moss protonema；E, picking moss protonema carry out the gametophyte that Liquid static culture obtains differentiation, and gametophyte is put into ventilation culture in fluid nutrient medium；F, by desert algae with fluid nutrient medium ventilation culture to exponential phase, obtain algae slurry after removing supernatant；G, liver moss gametophyte and desert algae algae slurry mixs after be seeded to prone soil surface, cultivate the compound skinning of acquisition liver moss and desert algae.Easy to implement the method, easy to operate, accelerate the During Succession of Desert Area Biological Soil Crusts, it is achieved that arid and semi-arid lands's desertification soil is quickly administered.

Description

A kind of liver moss is combined the breeding method of skinning with desert algae
Technical field
The invention belongs to field of environment engineering technology, is related to the restoration of the ecosystem of desertification soil, a kind of tongue is more particularly to Moss is combined the breeding method of skinning with desert algae.
Background technology
Biological Soil Crusts（Biological soil crusts）It is algae preferred growth, after native table is fixed, other Cryptogam subsequently also grows the topsoil for being formed thereon.They are distributed widely in global arid and semi-arid lands, Play carbon sequestration, fixed nitrogen, stable soil, change the life of spectral reflectance and moisture condition, impact vascular plant in these regions The important Ecology Actions such as long and seed sprouting.Biological Soil Crusts are generally made up of fungi, lichens, liver moss and algae.According to In Biological Soil Crusts, the biological difference of main composition, can be classified as algae skinning, lichens skinning and moss crust.Normal conditions Lower algae skinning is the primary stage that Biological Soil Crusts are formed, after only algae skinning is formed and stablizes topsoil, lichens, liver moss Be possible to plant a colony Deng cryptogam.And moss crust is the advanced stage that Biological Soil Crusts develop succession, bryophyte one After denier is planted a colony, accelerate the formation of soil by all means, and gradually improve the native liquid manure situation of table, when organic layer reaches foot During enough thickness, vascular plant just can be planted a colony.
The performance directly perceived of arid and semi-arid regions ecological degradation is exactly desertification of land.Although Desertification Soil is being eliminated Lentamente can recover after artificial disturbance, but by the natural deduction improvement table soil of Biological Soil Crusts so as to realize vascular plant Plant a colony, need the very long time.Because in its natural state, the development of Biological Soil Crusts is affected by various factors And restriction, its During Succession is very slow.The succession for hence speeding up Biological Soil Crusts has for the improvement of Desertification Soil Important meaning.
At present, the method for forming algae skinning in desertificated area using the desert algae of artificial infection large-scale culture has Perfect technical system and application practice, and there are some technical disadvantages in the exploration for being directed to moss crust breeding method.China Patent（Application）Number for 200410098476.x patent provide using bryophyte administer stony desertification method；Chinese patent （Application）Number for 201410586406.2 patent provide a kind of fast breeding method of loess plateau moss crust；China is specially Profit（Application）Number for 201410691489.1 patent provide a kind of fast breeding method of sand ground moss crust.Above-mentioned patent Cultivation to moss crust has carried out certain exploration, but its common ground is liver moss material depends on a large amount of collections in field, and This is not particularly suited for arid area and desertificated area.Chinese patent（Application）Number provide for the patent of 200810035554.x Desert muskeg and its method for biological breadcrust quick proliferation, the technology can be realized the interior of desert muskeg skinning and train on a small scale Support, and relatively difficult to achieve in large-scale production, while not being suitable for desertificated area yet.
In desertificated area, liver moss plant a colony be by algae skinning to the fixation of table soil and improvement based on, therefore in famine Moss crust is directly cultivated in desertization area is used for the necessary material base of desertification treatment shortage and environmental condition.
Content of the invention
In order to solve a difficult problem present in prior art, the purpose of the present invention is to there are provided a kind of liver moss and desert algae The breeding method of compound skinning, easy to implement the method, easy to operate, easy maintenance, accelerate drilling for Desert Area Biological Soil Crusts For process, it is achieved that the improvement rapidly and efficiently of arid and semi-arid lands's desertification soil.
The present invention is employed the following technical solutions：
A kind of liver moss is combined the breeding method of skinning with desert algae, and its step is：
1. desertification waits that administering regional liver moss sociales confirms the collection with moss crust and preservation：According to extensive investigations, In summer identification area natural habitat to be repaired, the liver moss dominant species of Biological Soil Crusts, select the well-developed tongue of the species Moss skinning, with cutting ring collection top layer skinning, room temperature（20-25 °C, same as below）Down air-dry 10-15 days standby.
2. the activation recovering of moss crust：Air-dried liver moss sample is sprayed water to cultivate at least 48 hours to after moisten completely and is made Its activation recovering；In liver moss sample activity recovery process, temperature setting is 10-15 °C, and intensity of illumination is 5-20 μm of ol m-2s-1, light dark period is 12/12, and humidity is 60%-80%.
3. the sandy soil that the moss crust after activation recovering includes are washed out with distilled water, obtain liver moss gametophyte.By obtained Liver moss gametophyte clean in sterile distilled water after disinfection.Disinfect with 5% hypochlorite disinfectant 10 of mass fraction Second, then three times are cleaned with aseptic distilled water again.
4. solid culture obtains protonema：Liver moss gametophyte after sterilization is put in solid medium to be cultivated to obtain Protonema is obtained, described solid medium is the BG11 solid medium containing mass fraction 1%-1.5% agar.Condition of culture has Body is：Temperature is respectively 25-30 °C of daytime, 10-15 °C of night, and light dark period is 12/12 hour, ambient humidity 60%-80%, Intensity of illumination is 15 μm of ol m of Initial stage of culture-2s-1, when starting amount reproduction after protonema adapts to environment, intensity of illumination increases For 70-100 μm of ol m-2s-1.Cultivation cycle is 15-20 days.
5. Liquid Culture obtains a large amount of gametophytes：The moss protonema that a large amount of picking solid culture are obtained under the microscope, Moss protonema is put into quiescent culture in CT fluid nutrient medium, cultivation cycle is 20-30 days, and moss protonema is divided into liver moss Gametophyte.Liver moss gametophyte is put in liquid CT culture medium carries out ventilation Amplification Culture, obtains the tongue that can be used for being inoculated with a large number Moss gametophyte, cultivation cycle are 30-40 days.Described CT fluid nutrient medium, its formula is：150 mg L-1Four water-calcium nitrate （Ca(NO3)2·4H2O）、100 mg L-1Potassium nitrate（KNO3）、40 mg L-1Epsom salt（MgSO4·7H2O）、25 mg L-1Five water β sodium glycero-phosphates（β-Na2glycerophosphate·5H2O）、400 mg L-1The third sulphur of Pehanorm base Acid（C7H17NO6S）、2.25 mg L-1Disodium ethylene diamine tetraacetate（EDTA-Na2）、0.123 mg L-1Tetrahydrate manganese chloride （MnCl2·4H2O）、0.015 mg L-1Seven water zinc chloride（ZnCl2·7H2O）、0.012 mg L-1Sodium Molybdate Dihydrate （Na2MoO4·2H2O）、0.291 mg L-1Iron trichloride hexahydrate（FeCl3·6H2O）、0.006 mg L-1CoCL2 6H2O （CoCl2·6H2O）.
6. desert algae algae kind and Liquid Culture obtain a large amount of living beings：Desert algae is cultivated extremely with fluid nutrient medium ventilation Exponential phase, obtains algae slurry after removing part supernatant.Described desert algae is Microccoleus（Microcoleus）, Scytonema arcangelii Category（Scytonema）, Nostoc（Nostoc）, Phormidium（Phormidium）, sheath Ulothrix（Lyngbya）One kind therein Or two to five kinds of any mixing, desert algae culture used medium is liquid B G11 culture medium.
7. combined inoculation, skinning maintenance and formation liver moss and desert algae are combined skinning：By liver moss gametophyte and desert algae algae Prone soil surface is seeded to after slurry mixing, and periodic maintenance, cultivation obtain the compound skinning of liver moss and desert algae.
In above-mentioned steps 4 and step 6, described BG11 culture medium prescription is：1500 mg L-1Sodium nitrate（NaNO3）、75 mg L-1Epsom salt（MgSO4·7H2O）、40 mg L-1Three water dipotassium hydrogen phosphates（K2HPO3·3H2O）、36 mg L-1 Calcium chloride dihydrate（CaCl2·2H2O）、20 mg L-1Sodium carbonate（Na2CO3）、6 mg L-1Citric acid（C6H8O7）、6 mg L-1 Ferric citrate（C6H10FeNO8）、1 mg L-1Disodium ethylene diamine tetraacetate（EDTA-Na2）、2.86 mg L-1Boric acid （H3BO3）、1.86 mg L-1Tetrahydrate manganese chloride（MnCl2·4H2O）、0.39 mg L-1Sodium Molybdate Dihydrate（Na2MoO4· 2H2O）、0.22 mg L-1White vitriol（ZnSO4·7H2O）、0.08 mg L-1Cupric sulfate pentahydrate（CuSO4·5H2O）、 0.05 mg L-1Cobalt nitrate hexahydrate（Co(NO3)2·6H2O）.
In above-mentioned steps 5 and step 6, described condition of culture is specially：Temperature is respectively 25-30 °C of daytime, night 10- 15 °C, light dark period is 12/12 hour, and intensity of illumination is 70-100 μm of ol m-2s-1.
In above-mentioned steps 5 and step 6, described ventilation culture is the gas by gas pumping liquid culture medium with air pump Body is filtered to air with Millipore Millex-GP filter before being passed through nutrient solution.
In above-mentioned steps 6, described nutrient solution natural subsidence removed part supernatant after 12 hours, obtained algae slurry.
In above-mentioned steps 7, liver moss gametophyte and desert algae（One or more mixing）With dry weight ratio as 1:2-1:10 ratio Mix and be inoculated with.
In above-mentioned steps 7, described periodic maintenance flow process is：It is inoculated with latter 15 days, was watered once with micro- spray per 6 hours, Every time 10-20 minute, discharge is 30 mL m-2min-1, watered once per 12 hours after 15 days, stop watering after 30 days；? Optionally apply the BG11 nutrient solution after 1-2 dilution during this.
The present invention compared with prior art, with advantages below and effect：
1., using the invention inoculation liver moss and desert algae, liver moss and desert algae composite junction can be formed on prone soil surface quickly Skin, Desert algal crust are slowly improved to topsoil environment after being formed, and are the planting a colony there is provided necessary of moss crust Material base, accelerates the succession process of skinning.
2. liver moss gametophyte can be obtained in a large number with fluid nutrient medium culture moss protonema, solve in prior art Liver moss material relies on the problem that field gathers in a large number mostly.
3. liver moss and the cultivation of the compound skinning of desert algae, improve species diversity and the life of artificial bio-membrane's soil crust Deposit ability so as to various matrix conditions and environment can be better adapted to.
Description of the drawings
Fig. 1 is the breeding method flow chart that a kind of liver moss and desert algae are combined skinning.
Wherein：1- desertification is waited to administer the confirmation of regional liver moss sociales, the collection of 2- moss crust and preservation, 3- liver moss Skinning activation recovering, 4- solid culture obtain protonema, 5- Liquid Culture obtain a large amount of gametophytes, 6- desert algae algae kind, 7- Liquid Culture obtains a large amount of living beings, 8- combined inoculation, the maintenance of 9- skinning, 10- formation liver moss and desert algae composite junction Skin.
Fig. 2 is that a kind of culture of true moss protonema gametophytic differentiation schematic diagram in a large number occurs after 20 days.
In figure filament shape is true moss protonema, and remaining is gametophyte.
Fig. 3 obtains a large amount of liver moss gametophyte schematic diagrames when being a kind of true moss protonema Liquid Culture 60 days.
Specific embodiment
Demonstration explanation is carried out to this inoculation technique below in conjunction with specific embodiment.
Embodiment 1：
According to Fig. 1, a kind of liver moss is combined the breeding method of skinning with desert algae, and its step is：
1. desertification is waited to administer collection and the preservation 2 that regional liver moss sociales confirm 1 and moss crust：According to extensive investigations, In summer identification area natural habitat to be repaired, the liver moss dominant species of Biological Soil Crusts, select the species well-developed Moss crust, with cutting ring collection top layer skinning, air-dry under room temperature 10 or 13 or 15 days standby.
2. moss crust activation recovering 3：The activation recovering of moss crust, air-dried liver moss sample are sprayed water to moistening completely After cultivate at least 48 hours and make its activation recovering；In liver moss sample activity recovery process, temperature setting is 10 or 12 or 14 or 15 ° C, intensity of illumination are 5 or 10 or 15 or 20 μm of ol m-2s-1, light dark period is 12/12, and humidity is 60 or 65 or 70 or 80%.
3. the sandy soil that the moss crust after activation recovering includes are washed out with distilled water, obtain liver moss gametophyte.By obtained Liver moss gametophyte clean in sterile distilled water after disinfection.Disinfect with 5% hypochlorite disinfectant 10 of mass fraction Second, then three times are cleaned with aseptic distilled water again.
4. solid culture obtains protonema 4：Gametophyte after sterilization is put in solid medium to be cultivated to obtain original Filament.Described solid medium is the BG11 solid medium containing mass fraction 1 or 1.2 or 1.4 or 1.5% agar.Culture Condition is specially：Temperature is respectively daytime 25 or 28 or 30 °C, night 10 or 13 or 15 °C, and light dark period is 12/12 hour, Ambient humidity 60 or 65 or 70 or 75 or 80%, intensity of illumination are 15 μm of ol m of Initial stage of culture-2s-1, after protonema adapts to environment When starting amount reproduction, it is 70 or 80 or 90 or 100 μm of ol m that intensity of illumination increases-2s-1.Cultivation cycle is 15 or 17 or 20 My god.
5. Liquid Culture obtains a large amount of gametophytes 5：The liver moss precursor that a large amount of picking solid culture are obtained under the microscope Body, moss protonema is put into quiescent culture in CT fluid nutrient medium, and cultivation cycle is 20-30 days, and moss protonema is divided into Liver moss gametophyte.Liver moss gametophyte is put in liquid CT culture medium carries out ventilation Amplification Culture, obtains and can be used in a large number being inoculated with Liver moss gametophyte, cultivation cycle be 30-40 days.Described CT fluid nutrient medium, its formula is：150 mg L-1Four water nitric acid Calcium（Ca(NO3)2·4H2O）、100 mg L-1Potassium nitrate（KNO3）、40 mg L-1Epsom salt（MgSO4·7H2O）、25 mg L-1Five water β sodium glycero-phosphates（β-Na2glycerophosphate·5H2O）、400 mg L-1Pehanorm base Propane sulfonic acid（C7H17NO6S）、2.25 mg L-1Disodium ethylene diamine tetraacetate（EDTA-Na2）、0.123 mg L-1Tetrahydrate manganese chloride （MnCl2·4H2O）、0.015 mg L-1Seven water zinc chloride（ZnCl2·7H2O）、0.012 mg L-1Sodium Molybdate Dihydrate （Na2MoO4·2H2O）、0.291 mg L-1Iron trichloride hexahydrate（FeCl3·6H2O）、0.006 mg L-1CoCL2 6H2O （CoCl2·6H2O）.
6. desert algae algae kind 6 and Liquid Culture obtain a large amount of living beings 7：By desert algae fluid nutrient medium ventilation culture To exponential phase, algae slurry after removing part supernatant, is obtained.Described desert algae is Microccoleus（Microcoleus）, pseudo- branch Trentepohlia（Scytonema）, Nostoc（Nostoc）, Phormidium（Phormidium）, sheath Ulothrix（Lyngbya）Therein one Kind or two to five kinds of any mixing, desert algae culture used medium is liquid B G11 culture medium.
7. combined inoculation 8, skinning maintenance 9 and formation liver moss and desert algae are combined skinning 10：By liver moss gametophyte and desert Prone soil surface is seeded to after the slurry mixing of algae algae, periodic maintenance, cultivation obtain the compound skinning of liver moss and desert algae.
In above-mentioned steps 4 and step 6, described BG11 culture medium prescription is：1500 mg L-1Sodium nitrate（NaNO3）、75 mg L-1Epsom salt（MgSO4·7H2O）、40 mg L-1Three water dipotassium hydrogen phosphates（K2HPO3·3H2O）、36 mg L-1 Calcium chloride dihydrate（CaCl2·2H2O）、20 mg L-1Sodium carbonate（Na2CO3）、6 mg L-1Citric acid（C6H8O7）、6 mg L-1 Ferric citrate（C6H10FeNO8）、1 mg L-1Disodium ethylene diamine tetraacetate（EDTA-Na2）、2.86 mg L-1Boric acid （H3BO3）、1.86 mg L-1Tetrahydrate manganese chloride（MnCl2·4H2O）、0.39 mg L-1Sodium Molybdate Dihydrate（Na2MoO4· 2H2O）、0.22 mg L-1White vitriol（ZnSO4·7H2O）、0.08 mg L-1Cupric sulfate pentahydrate（CuSO4·5H2O）、 0.05 mg L-1Cobalt nitrate hexahydrate（Co(NO3)2·6H2O）.
In above-mentioned steps 5 and step 6, described condition of culture is specially：Temperature is respectively daytime 25 or 27 or 30 °C, black Night 10 or 13 or 15 °C, light dark period are 12/12 hour, and intensity of illumination is 70 or 80 or 90 or 100 μm of ol m-2s-1.
In above-mentioned steps 5 and step 6, described air pump carries out ventilation culture, gas be passed through nutrient solution before with Millipore Millex-GP filter is filtered to air.
In above-mentioned steps 6, described nutrient solution natural subsidence removed part supernatant after 12 hours, obtained algae slurry.
In above-mentioned steps 7, liver moss gametophyte and desert algae（One or more mixing）With dry weight ratio as 1:2-1:10 ratio Mix and be inoculated with, the liver moss gametophyte of inoculation is 0.3-1.0 g m with desert algae gross dry weight-2.
In above-mentioned steps 7, described periodic maintenance flow process is：It is inoculated with latter 15 days, was watered once with micro- spray per 6 hours, 10 or 13 or 15 or 18 or 20 minutes every time, discharge was 30 mL m-2min-1, watered once per 12 hours after 15 days, 30 days Stop afterwards watering；Optionally apply the BG11 nutrient solution after 1-2 dilution in the process.
The initial composite skinning of true moss and the micro- sheath algae of tool sheath can be formed after safeguarding a period of time.
Embodiment 2：
According to Fig. 1, a kind of liver moss is combined the breeding method of skinning with desert algae, and its step is：
Investigation finds true moss（Bryum argenteum）With autochthonal to tooth moss（Didymodon vinealis）In being distributed widely in Mongolian autonomous region Dalate Banner.Well-developed two kinds of moss crust are selected, is preserved after air-drying respectively.Air-dried true moss sample spray Water, to after moisten completely, is 15 °C in temperature, and intensity of illumination is 20 μm of ol m-2s-1, light dark period is 12/12, and humidity is Cultivating under conditions of 80% at least 48 hours makes its activation recovering.True moss skinning distilled water after activity recovery washes out sandy soil, obtains Obtain true moss gametophyte.Gametophyte is cleaned three times in sterile distilled water.Subsequently 5% hypochlorite disinfectant of mass fraction is used 10 seconds Clock, is then cleaned with distilled water three times again.True moss gametophyte after sterilization is positioned on BG11 solid medium and is cultivated.Culture Condition is：Temperature is respectively 30 °C of daytime, 15 °C of night, and light dark period is 12/12 hour, humidity 80%, and intensity of illumination is 70 μmol m-2s-1.Cultivation cycle is 18 days.The moss protonema that a large amount of picking solid culture are obtained under the microscope, liver moss is former Filament is put into quiescent culture in CT fluid nutrient medium, and cultivation cycle is 20-30 days, and moss protonema is divided into liver moss gametophyte. Condition of culture is：Temperature is respectively 30 °C of daytime, 15 °C of night, and light dark period is 12/12 hour, and intensity of illumination is first for culture 15 μm of ol m of phase-2s-1, when starting amount reproduction after protonema adapts to environment, intensity of illumination increases to 100 μm of ol m-2s-1. There is gametophytic differentiation in a large number after 20 days in culture（Fig. 2）.Liver moss gametophyte is put in liquid CT culture medium carries out ventilation expansion Big culture, obtains the liver moss gametophyte that can be used for being inoculated with a large number, and cultivation cycle is 30-40 days.Condition of culture is：Temperature is respectively 30 °C of daytime, 15 °C of night, light dark period are 12/12 hour, and intensity of illumination is 100 μm of ol m-2s-1.With incubation time Increase, still in lasting growth, culture has been almost full with entirely the gametophytic quantity of true moss to true moss gametophyte when 60 days Conical flask（Fig. 3）.
With the micro- sheath algae of BG11 culture medium ventilation culture tool sheath in the serum bottle of 10 L（Microcoleus vaginatus）, condition of culture is：Temperature is respectively 30 °C of daytime, 15 °C of night, and light dark period is 12/12 hour, and illumination is strong Spend for 100 μm of ol m-2s-1.After cultivating to exponential phase, natural subsidence removes part supernatant and obtains algae slurry after 12 hours.
The gametophyte of liver moss and the micro- sheath algae of tool sheath are 1 according to dry weight:8 ratio is mixed, and is sprayed after mixing Sand surface is inoculated in, the liver moss gametophyte of inoculation is 0.9 g m with the micro- sheath algae gross dry weight of tool sheath-2.
It is inoculated with latter 15 days, was watered once with micro- spray per 6 hours, each 10-20 minute, discharge is 30 mL m-2 min-1, watered once per 12 hours after 15 days, stop watering after 30 days；After optionally applying 1-2 dilution in the process BG11 nutrient solution.
The initial composite skinning of true moss and the micro- sheath algae of tool sheath can be formed after one month, and it is left that bio-crust coverage can reach 90% The cover degree on the right side, wherein liver moss can reach 60%-90%.
Other implementation steps are same as Example 1.
Embodiment 3：
Autochthonal to the gametophytic training method of tooth moss with embodiment 1.
Culture has the micro- sheath algae of sheath and scytonema javanicum respectively（Scytonema javanicum）To logarithmic phase, training method With embodiment 1 or embodiment 2.
By autochthonal be 1 with the micro- sheath algae of tool sheath and scytonema javanicum according to dry weight to tooth moss gametophyte:5:3 ratio is mixed Close, sprinkling after mixing is inoculated in sand surface, and the liver moss gametophyte of inoculation is always done with the micro- sheath algae of tool sheath and scytonema javanicum Weight is 0.9 g m-2.
Skinning is postvaccinal to be safeguarded with embodiment 1 or embodiment 2.
Can be formed after one month autochthonal to tooth moss and the micro- sheath algae of tool sheath and the initial composite skinning of scytonema javanicum, skinning Cover degree can reach 94% or so, and the wherein cover degree of liver moss can reach 50%-92%.
Specific embodiment described herein is only explanation for example to present invention spirit.The affiliated technology neck of the present invention The technical staff in domain can be made various modifications or supplement or replaced using similar mode to described specific embodiment Generation, but the spirit without departing from the present invention or surmount scope defined in appended claims.

Claims (5)

1. a kind of liver moss and desert algae are combined the breeding method of skinning, and its step is：
A, desertification are waited to administer the confirmation of regional liver moss sociales and collection and the preservation of moss crust：According to extensive investigations, The advantage liver moss species of Biological Soil Crusts in summer identification area natural habitat to be repaired, selects the liver moss knot of the species development Skin, with cutting ring collection top layer skinning, air-dry under room temperature 10-15 days standby；
B, the activation recovering of moss crust：Air-dried liver moss sample sprays water to cultivate at least 48 hours to after moisten completely makes its activity Recover；In liver moss sample activity recovery process, temperature setting is 10-15 °C, and intensity of illumination is 5-20 μm of ol m-2s-1, brightness Cycle is 12/12, and humidity is 60%-80%；
C, the sandy soil that the moss crust of activation recovering includes are washed out with distilled water, obtain liver moss gametophyte, the liver moss of acquisition is joined Daughter clean in sterile distilled water after disinfection, disinfect the hypochlorite disinfectant 10 seconds with mass fraction 5%, Three times are cleaned again with aseptic distilled water；
D, solid culture obtain protonema：Liver moss gametophyte after sterilization is put in solid medium to be cultivated to obtain precursor Body, described solid medium are the BG11 solid medium containing mass fraction 1%-1.5% agar, and condition of culture is：Temperature Respectively 25-30 °C of daytime, 10-15 °C of night, light dark period is 12/12 hour, ambient humidity 60%-80%, and intensity of illumination is 15 μm of ol m of Initial stage of culture-2s-1, when starting amount reproduction after protonema adapts to environment, intensity of illumination increases as 70-100 μ mol m-2s-1, cultivation cycle is 15-20 days；
E, Liquid Culture obtain gametophyte：The moss protonema that picking solid culture is obtained under the microscope, by moss protonema Quiescent culture in CT fluid nutrient medium is put into, cultivation cycle is 20-30 days, and moss protonema is divided into liver moss gametophyte, by tongue Moss gametophyte is put in liquid CT culture medium and carries out ventilation Amplification Culture, obtains the liver moss gametophyte for being inoculated with, cultivation cycle For 30-40 days, described CT fluid nutrient medium, its formula is：150 mg L-1Four water-calcium nitrate, 100 mg L-1Potassium nitrate, 40 mg L-1Epsom salt, 25 mg L-1Five water β sodium glycero-phosphates, 400 mg L-1Pehanorm base propane sulfonic acid, 2.25 mg L-1Disodium ethylene diamine tetraacetate, 0.123 mg L-1Tetrahydrate manganese chloride, 0.015 mg L-1Seven water zinc chloride, 0.012 mg L-1Sodium Molybdate Dihydrate, 0.291 mg L-1Iron trichloride hexahydrate, 0.006 mg L-1CoCL2 6H2O；
F, desert algae algae kind and Liquid Culture obtain a large amount of living beings：Desert algae is given birth to logarithm with fluid nutrient medium ventilation culture For a long time, algae slurry is obtained after removing supernatant, and described desert algae is Microccoleus, Scytonema, Nostoc, Phormidium, sheath silk Therein a kind of or two to five kinds of any mixing of Trentepohlia, desert algae culture used medium are liquid B G11 culture medium；
G, combined inoculation, skinning maintenance and formation liver moss and desert algae are combined skinning：Will be mixed with desert algae algae slurry for liver moss gametophyte Prone soil surface is seeded to after conjunction, and periodic maintenance, cultivation obtain the compound skinning of liver moss and desert algae.
2. a kind of liver moss according to claim 1 and desert algae are combined the breeding method of skinning, it is characterised in that：Described BG11 culture medium prescription is：1500 mg L-1Sodium nitrate, 75 mg L-1Epsom salt, 40 mg L-1Three water phosphoric acid hydrogen two Potassium, 36 mg L-1Calcium chloride dihydrate, 20 mg L-1Sodium carbonate, 6 mg L-1Citric acid, 6 mg L-1Ferric citrate, 1 mg L-1Disodium ethylene diamine tetraacetate, 2.86 mg L-1Boric acid, 1.86 mg L-1Tetrahydrate manganese chloride, 0.39 mg L-1Two water Sodium molybdate, 0.22 mg L-1White vitriol, 0.08 mg L-1Cupric sulfate pentahydrate, 0.05 mg L-1Cobalt nitrate hexahydrate.
3. a kind of liver moss according to claim 1 and desert algae are combined the breeding method of skinning, it is characterised in that：Described The condition of culture of step E and F is：Temperature is respectively 25-30 °C of daytime, 10-15 °C of night, and light dark period is 12/12 hour, Intensity of illumination is 70-100 μm of ol m-2s-1.
4. a kind of liver moss according to claim 1 and desert algae are combined the breeding method of skinning, it is characterised in that：Described Liver moss gametophyte is with desert algae with dry weight ratio as 1:2-1:10 ratio is mixed and is inoculated with.
5. a kind of liver moss according to claim 1 and desert algae are combined the breeding method of skinning, it is characterised in that：Described Periodic maintenance flow process is：It is inoculated with latter 15 days, was watered once with micro- spray per 6 hours, each 10-20 minute, discharge is 30 mL m-2min-1, watered once per 12 hours after 15 days, stop watering after 30 days；Optionally apply 1-2 dilution in the process BG11 nutrient solution afterwards.
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CN108782089A (en) * 2018-05-04 2018-11-13 永嘉县原野园林工程有限公司 A kind of breeding method of moss
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CN111264370A (en) * 2020-01-17 2020-06-12 中山大学 Production system and production method for repairing ecological blanket in tailing pond
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CN113906854A (en) * 2021-10-08 2022-01-11 中国科学院西北生态环境资源研究院 Method for treating saline-alkali soil in arid region by utilizing artificial composite biological crust
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CN114365686B (en) * 2022-01-07 2022-12-09 西南科技大学 Method for cultivating moss for biological monitoring
CN114752392A (en) * 2022-04-26 2022-07-15 北京林业大学 Method for promoting artificial rapid cultivation of biological crust by functional microbial agent
CN114752392B (en) * 2022-04-26 2024-03-29 北京林业大学 Method for promoting artificial rapid cultivation of biological crust by using functional microbial agent

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